Method for screening veneridae mitochondria COI gene amplification primers

Abstract

The invention discloses a method for screening veneridae mitochondria COI (cytochrome c oxidase subunit I) gene amplification primers. The method is characterized by comprising the following steps of: a, logging in National Center for Biotechnology Information, and downloading complete mitochondria sequences of veneridae shellfishes; b, comparing and analyzing the downloaded sequences by using BioEdit, and determining a conserved region of the sequences; c, designing a plurality of pairs of primers by using Primer Premier 5 in the conserved region and synthesizing the primers; d, extracting veneridae shellfish DNA, and performing PCR amplification on the synthesized primers under a corresponding reaction system; and 3, performing electrophoresis detection, and screening a group of cocktail primers with single stripe and high amplification efficiency, wherein the group of cocktail primers consists of a pair of degenerate primers and a pair of common primers. The veneridae mitochondria COI primers can effectively amplify destination fragments of different species of the veneridae, and have significance for COI sequence-based veneridae species classification, phylogenesis, systematicgeography research, veneridae germ plasm resource protection and fishery resource management.

Description

Technical field [0001] The present invention relates to a method for screening mitochondrial cytochrome c oxidase subunit I (COI) gene sequence amplification primers of bivalve mollusks, in particular to a mitochondrial COI gene sequence amplification The screening method of primers. Background technique [0002] Veneridae (Veneridae) shellfish belong to Mollusca, Biavalvia, Heterodonta, Veneroida, Veneracea, which are There are shellfish groups distributed all over the world, large in number, many economic types, and high commercial value. According to statistics, there are more than 800 species of shellfish in the clam family worldwide, belonging to 14 subfamilies, and the most abundant family of bivalves. Clam shellfish contain many species with high economic value, such as Ruditapesphilippinarum, Gomphina aequilatera, Saxidomus purpuratus, Meretrix meretrix, Cyclina sinensis, etc. However, many species of Clamaceae have great morphological plasticity, and the morphological...

AI Technical Summary

Problems solved by technology

[0004] However, the variation rate of shellfish COI sequences is significantly higher than that of other groups of animals: different species of molluscs, even close relatives and even different groups within a species, have great variation in COI sequences, which led to Folmer et al. 1994 The amplification efficiency of the COI universal primers LCO1490 and HCO2198 designed in 2009 was extremely low in this animal group

The lack of primers with high amplification efficiency has seriously hindered the research on the phylogeny of clam shellfish